PROJECT SUMMARY/ABSTRACT The Redox Metabolism Shared Resource Facility (RM SRF) is a cancer-center managed SRF that provides critical services for University of Kentucky (UK) Markey Cancer Center (MCC) Research Programs. Notably, the RM SRF is one of only a small number of facilities directly associated with a National Cancer Institute-designated Cancer Center that provides expertise and analyses in oxidative stress, cellular energetics, metabolomics and proteomics in cancer and cancer biology. Over the last 4 years of CCSG funding, the RM SRF has successfully demonstrated its high value-added role for MCC investigators doing basic science, clinical and cancer prevention and control research in the MCC's Kentucky catchment area. There is increasing evidence that oxidative damage, with sequelae of altered cellular energetics and metabolism as well as protein alterations, underlie the initiation, promotion, invasion, metastasis and treatment of cancers. Moreover, oxidative stress is implicated in the side effects of normal tissue injury following cancer therapy. As such, the 4 basic services provided by the RM SRF are: 1) analysis of markers of oxidative and nitrosative stress; 2) molecular biological manipulation of biological systems to investigate redox signals, including measurements of mitochondrial function by Seahorse technology; 3) proteomics identification of differentially expressed, differentially oxidized or differentially covalently modified proteins in various cancer- related systems; and 4) profiling and Stable Isotope Resolved Metabolomics of cancer tissue and cells, a service that began in 2015 following a $6M UK investment in nuclear magnetic resonance and mass spectrometry equipment. Key improvements since initial CCSG funding include expanding services and scope toward metabolomics as well as increasing staff. This SRF is directed by Dr. D. Allan Butterfield (GEM) with Dr. Haining Zhu (GEM), as Associate Director. With the exciting addition of metabolomics analyses to the SRF, Dr. Richard Higashi (GEM), an expert in the assessment of metabolites and metabolic pathways, has been added as Associate Director of RM SRF for metabolomics. Fifty-four MCC members used RM SRF services since MCC funding began, a 43% increase from year 1 to year 4 of CCSG funding. Of these, 39 were members with peer-reviewed funding. RM SRF leadership and staff actively participate in the development of new applications and methodologies for a better understanding of the roles of oxidative stress and metabolism in cancer and cancer chemotherapy and provide technical assistance with experimental design and interpretation of results for MCC principal investigators. RM SRF technical staff have knowledge of the latest and most current sample handling and preparation methods so that reliable, precise and artifact-free assay results can be obtained. The RM SRF facilitates research with the other MCC SRFs to improve the analytical power of various cutting-edge technologies for understanding of the enormous cancer burden in Kentucky.